CN101073666B - Method for producing kallidin proenzyme raw-material medicine - Google Patents
Method for producing kallidin proenzyme raw-material medicine Download PDFInfo
- Publication number
- CN101073666B CN101073666B CN2007100982895A CN200710098289A CN101073666B CN 101073666 B CN101073666 B CN 101073666B CN 2007100982895 A CN2007100982895 A CN 2007100982895A CN 200710098289 A CN200710098289 A CN 200710098289A CN 101073666 B CN101073666 B CN 101073666B
- Authority
- CN
- China
- Prior art keywords
- liquid
- kallidin
- proenzyme
- raw
- material medicine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Enzymes And Modification Thereof (AREA)
Abstract
The invention is concerned with the method for prepareing high purity organism pharmaceutical product, that is, the method of the high purity pancreatic kiniogenase pharmaceutical product. The method includes: isolates the pancreatic kinigenase midbody perfectly, passes it through the QAE-Sepharose Fast Flow that generates the pharamaceutical prodct of the kinigenase by hydronium column chromatographic purification. This pharamaceutical produc approaches high level of purity with 75%, increases the titer to 55 unit/ mg and above, and boots up the specific activity to 350 unit/mg albumen and above.
Description
Technical field
The present invention relates to a kind of method for preparing high-purity biological raw material medicine, particularly a kind of method for preparing high-purity kallidin proenzyme raw-material medicine.
Background technology
Kallidinogenase is a kind of peripheral vasodilators with the effect of blood vessel dilating microcirculation improvement.It can activate fibrinolysin, inhibition of phospholipase A
2, have the reduction blood viscosity, prevent platelet aggregation, prevent effects such as thrombosis.It is mainly used in diseases with microcirculatory disturbance, and the treatment of nephropathy, peripheral neuropathy, retinopathy, retinopathy and the ischemic cerebrovascular that causes as diabetes also can be used for the auxiliary treatment of hypertension.
Kallidinogenase obtains through extracting purification from mammiferous pancreas, promptly extracts from pancreas itself, pancreas enzyme powder and kallidinogenase crude product (intermediate), and makes by purification.The purification process of existing kallidinogenase comprises that use polysaccharide non-resin material carries out purification, for example makes thick kallidinogenase be adsorbed onto on a kind of weakly basic anion exchange fibre element and separation (seeing Japan Patent distribution No.11697/62); Perhaps a kind of aluminum salt or zinc salt adding are contained in the aqueous solution of kallidinogenase, be adsorbed onto the precipitation of the kallidinogenase that produces on the weakly alkaline cellulose anion exchanger or cross-linking dextran (Sephadex) on, and separate it (seeing Japanese publication distribution No.56886/73); Another kind of known purification process is to use ion exchange resin, promptly add a kind of protein precipitant in the pancreas extracting solution, the kallidinogenase that produces is adsorbed onto the macroporous strong basic anion exchange resin, and separates it (seeing Japanese publication No.103715/73).Yet utilizing method shortcoming that above-mentioned glycan non-resin material adsorbs is that the physical strength of ion exchange material is not enough, can not carry out mass preparation.And ion-exchange-resin process can not make product remove unwanted impurity when eluting.Therefore, the said method purity that obtains product awaits further to improve.
" Chinese biochemical drug magazine " 2005 Vol.26 No.1P.37-38 " ion exchange chromatography prepares high-purity pancreatic kininogenase " author's Zhang Jianxins, Rhizoma Zingiberis Recens mark, Liu Changliang.
The original method that adopts of inventor is by ion exchange chromatography single step purification kallidinogenase crude product, adopts 3-4 times of acetone precipitation, the kallidinogenase that purification obtains, and than living more than 300U/mg protein, activity recovery is more than 65%.But the price of original PK eluent is high, and can cause a large amount of losses in using the acetone precipitation process.And purity is not high yet.In order to solve above variety of problems, the inventor has adopted new process.
Summary of the invention
The object of the present invention is to provide a kind of method for preparing high-purity kallidin proenzyme raw-material medicine, this method under the separation condition of optimizing, is passed through the kininogenase raw material medicine that QAE-SepharoseFast Flow ion column chromatography purification makes by the kallidinogenase intermediate.The purity of this crude drug is up to more than 75%, and tiring increases by 55 units/more than the mg, reaches 350 units/more than the mg albumen, referring to table 1 and table 2 than living.
Method of the present invention is as follows:
1. preparation: the NH of preparation 0.15-0.5mol/L
4The NH of AC buffer and 0.1-0.5mol/L
4The AC buffer is adjusted to pH6.0, and surveys electric conductivity value;
2. chromatography:
2.1 balance: the chromatographic column of QAE-Sepharose Fast Flow ion column filler will be housed, use the NH of the 0.15-0.5mol/L of pH6.0
4AC buffer balance finishes until the solution pH value unanimity, the identical back balance of electric conductivity value that pass in and out chromatographic column;
2.2 sample introduction: transfer the pH value close with balance liquid (6.1～6.2) of kallidinogenase intermediate, electric conductivity value is a little less than balance liquid electric conductivity value (lower slightly 0.2～0.3 * 10
4μ s/cm), calculate applied sample amount according to every liter of filler maximum exchange amount 2,000 ten thousand units, with the sample solution upper prop;
2.3 washing: the NH that uses the 0.15-0.5mol/L of pH6.0
4AC buffer flushing pillar is followed the tracks of with spectrophotometer simultaneously and is detected the absorption value of effluent at the 280nm place, washes to be lower than at 0.5 o'clock to the OD value of eluate and to stop;
2.4 eluting: the NH that uses the 0.1-0.5mol/L of pH6.0
4AC buffer solution elution pillar is followed the tracks of with spectrophotometer simultaneously and is detected its absorption value at the 253nm place, collects peak liquid;
3. ultrafiltration desalination: will collect liquid and add the purified water desalination and concentration by ultrafiltration, when exceeding the liquid electric conductivity value less than 0.1 * 10
4During μ s/cm, can stop to add purified water, continue to concentrate;
4. concentrated solution filters;
5. add the adjuvant lyophilizing;
6. sieve, packing.
The following condition of preferred use is operated in said method:
In step 1, the NH of preparation 0.15mol/L
4The NH of AC buffer and 0.5mol/L
4Adjust pH with acetic acid and ammonia respectively during the AC buffer.
In step 2, used chromatographic column is ￠ 40cm, and volume is 50L; Be that eluting is collected starting point when tiring greater than 50 units/cm with eluent, tiring less than 50 units/cm with eluent is that eluting is collected terminal point, collects the long-pending 30～60L that is about of peak liquid.
In step 3, the molecular cut off of ultrafiltration desalination is 10000dt; When being concentrated into liquid volume 6～11L, close ultrafilter, purified water is towards film, tire when being lower than 50 units/ml to return pipe, concentrated solution, the sampling censorship.
In step 4, the cartridge filter of concentrated solution with 0.22 μ m film filtered, get filtering and concentrating liquid.
In step 5, it is as follows to add the freeze dried concrete steps of adjuvant: according to tiring of concentrated solution, the control finished product than vigor 350 units/mg albumen above with tire in 70 units/more than the mg, calculate the consumption of gelatin, lactose, respectively with the water for injection heating for dissolving and by 0.22 μ m membrane filtration, to be cooled, add in the filtering and concentrating liquid, mix laggard freeze drying box lyophilizing.The freeze-dry process condition is: pre-freeze: products temperature-20 ℃--40 ℃, liquid measure in the optic disc is incubated 2-4 hour.Distillation: 10 ℃/hour of conduction oil programming rates, to oily temperature rise to 30 ℃, case vacuum is not more than 15Pa before the process control.Dry: as when products temperature reaches 23 ℃, to be incubated more than 4 hours until dry terminal point.
In step 6, finished product that lyophilizing is good is pulverized and is crossed behind 80 mesh sieves and the mix homogeneously with medicinal complex pocket packing, is loaded in aluminum listens, and seals.
The specific embodiment
Embodiment 1
1. preparation eluent
The NH of preparation 0.15mol/L
4The NH of AC buffer and 0.5mol/L
4The AC buffer is adjusted to pH6.0 with acetic acid and ammonia respectively, and surveys electric conductivity value.
2. chromatography
2.1 dress column equilibration: ion column filler QAE-Sepharose Fast Flow is loaded on ￠ 40cm, and volume is in the chromatographic column of 50L, uses the NH of the 0.15mol/L of pH6.0
4AC buffer balance finishes until turnover solution pH value unanimity, the identical back balance of electric conductivity value.
2.2 sample introduction: transfer kallidinogenase intermediate pH value close with balance liquid (6.1～6.2), electric conductivity value is a little less than balance liquid electric conductivity value (lower slightly 0.2～0.3 * 10
4μ s/cm), according to every liter of filler maximum exchange amount 2,000 ten thousand units, calculate applied sample amount.With the sample solution upper prop.
2.3 washing: the NH that uses the 0.5mol/L of pH6.0
4AC buffer flushing pillar is followed the tracks of with spectrophotometer simultaneously and is detected the absorption value of effluent at the 280nm place, washes to be lower than at 0.5 o'clock to eluate OD value and to stop.
2.4 eluting: the NH that uses the 0.5mol/L of pH6.0
4AC buffer solution elution pillar is followed the tracks of with spectrophotometer simultaneously and is detected its absorption value at the 253nm place, collects peak liquid.
3. ultrafiltration desalination
Add the purified water desalination and concentration by ultrafiltration 3.1 will collect liquid, when exceeding the liquid electric conductivity value less than 0.1 * 10
4When μ s/cm is following, can stop to add purified water, continue to concentrate.
3.2 when being concentrated into liquid volume 6～11L, close ultrafilter, purified water is towards film, tire when being lower than 50 units/ml to return pipe, concentrated solution, the sampling censorship.
4. concentrated solution filters: the cartridge filter of concentrated solution with 0.22 μ m film filtered, get filtering and concentrating liquid.
5. add the adjuvant lyophilizing: according to tiring of concentrated solution, the control finished product is above and tire in 70 units/more than the mg consumption of calculating gelatin, lactose than vigor 350 units/mg albumen.And respectively with the water for injection heating for dissolving and by 0.22 μ m membrane filtration, to be cooled to below 25 ℃, add the laggard freeze drying box lyophilizing of mix homogeneously in the filtering and concentrating liquid.The even packing 1～3L of every dish liquid is decided lyophilizing during lyophilizing on total amount of liquid.The freeze-dry process condition is: pre-freeze: products temperature-20 ℃--40 ℃, liquid measure in the optic disc is incubated 2-4 hour.Distillation: 10 ℃/hour of conduction oil programming rates, to oily temperature rise to 30 ℃, case vacuum is not more than 15Pa before the process control.Dry: as when products temperature reaches 23 ℃, to be incubated more than 4 hours until dry terminal point.
6. sieve, packing: finished product that lyophilizing is good is pulverized and is crossed behind 80 mesh sieves and the mix homogeneously with medicinal complex pocket packing, is loaded in aluminum listens, and seals.
Embodiment 2
1. preparation eluent
The NH of preparation 0.5mol/L
4The NH of AC buffer and 0.1mol/L
4The AC buffer is adjusted to pH6.0 with acetic acid and ammonia respectively, and surveys electric conductivity value.
2. chromatography
2.1 dress column equilibration: ion column filler QAE-Sepharose Fast Flow is loaded on ￠ 40cm, and volume is in the chromatographic column of 50L, uses the NH of the 0.15mol/L of pH6.0
4AC buffer balance finishes until turnover solution pH value unanimity, the identical back balance of electric conductivity value.
2.2 sample introduction: transfer kallidinogenase intermediate pH value close with balance liquid (6.1～6.2), electric conductivity value is a little less than balance liquid electric conductivity value (lower slightly 0.2～0.3 * 10
4μ s/cm), according to every liter of filler maximum exchange amount 2,000 ten thousand units, calculate applied sample amount.With the sample solution upper prop.
2.3 washing: the NH that uses the 0.5mol/L of pH6.0
4AC buffer flushing pillar is followed the tracks of with spectrophotometer simultaneously and is detected the absorption value of effluent at the 280nm place, washes to be lower than at 0.5 o'clock to eluate OD value and to stop.
2.4 eluting: the NH that uses the 0.1mol/L of pH6.0
4AC buffer solution elution pillar is followed the tracks of with spectrophotometer simultaneously and is detected its absorption value at the 253nm place, collects peak liquid.
3. ultrafiltration desalination
Add the purified water desalination and concentration by ultrafiltration 3.1 will collect liquid, when exceeding the liquid electric conductivity value less than 0.1 * 10
4When μ s/cm is following, can stop to add purified water, continue to concentrate.
3.2 when being concentrated into liquid volume 6～11L, close ultrafilter, purified water is towards film, tire when being lower than 50 units/ml to return pipe, concentrated solution, the sampling censorship.
4. concentrated solution filters: the cartridge filter of concentrated solution with 0.22 μ m film filtered, get filtering and concentrating liquid.
5. add the adjuvant lyophilizing: according to tiring of concentrated solution, the control finished product is above and tire in 70 units/more than the mg consumption of calculating gelatin, lactose than vigor 350 units/mg albumen.And respectively with the water for injection heating for dissolving and by 0.22 μ m membrane filtration, to be cooled to below 25 ℃, add the laggard freeze drying box lyophilizing of mix homogeneously in the filtering and concentrating liquid.The even packing 1～3L of every dish liquid is decided lyophilizing during lyophilizing on total amount of liquid.The freeze-dry process condition is: pre-freeze: products temperature-20---40 ℃, liquid measure in the optic disc is incubated 2-4 hour.Distillation: 10 ℃/hour of conduction oil programming rates, to oily temperature rise to 30 ℃, case vacuum is not more than 15Pa before the process control.Dry: as when products temperature reaches 23 ℃, to be incubated more than 4 hours until dry terminal point.
6. sieve, packing: finished product that lyophilizing is good is pulverized and is crossed behind 80 mesh sieves and the mix homogeneously with medicinal complex pocket packing, is loaded in aluminum listens, and seals.
Embodiment 3
1. preparation eluent
The NH of preparation 0.35mol/L
4The NH of AC buffer and 0.3mol/L
4The AC buffer is adjusted to pH6.0 with acetic acid and ammonia respectively, and surveys electric conductivity value.
2. chromatography
2.1 dress column equilibration: ion column filler QAE-Sepharose Fast Flow is loaded on ￠ 40cm, and volume is in the chromatographic column of 50L, uses the NH of the 0.15mol/L of pH6.0
4AC buffer balance finishes until turnover solution pH value unanimity, the identical back balance of electric conductivity value.
2.2 sample introduction: transfer kallidinogenase intermediate pH value close with balance liquid (6.1～6.2), electric conductivity value is a little less than balance liquid electric conductivity value (lower slightly 0.2～0.3 * 10
4μ s/cm), according to every liter of filler maximum exchange amount 2,000 ten thousand units, calculate applied sample amount.With the sample solution upper prop.
2.3 washing: the NH that uses the 0.35mol/L of pH6.0
4AC buffer flushing pillar is followed the tracks of with spectrophotometer simultaneously and is detected the absorption value of effluent at the 280nm place, washes to be lower than at 0.5 o'clock to eluate OD value and to stop.
2.4 eluting: the NH that uses the 0.3mol/L of pH6.0
4AC buffer solution elution pillar is followed the tracks of with spectrophotometer simultaneously and is detected its absorption value at the 253nm place, collects peak liquid.
3. ultrafiltration desalination
Add the purified water desalination and concentration by ultrafiltration 3.1 will collect liquid, when exceeding the liquid electric conductivity value less than 0.1 * 10
4When μ s/cm is following, can stop to add purified water, continue to concentrate.
3.2 when being concentrated into liquid volume 10L, close ultrafilter, purified water is towards film, tire when being lower than 50 units/ml to return pipe, concentrated solution, the sampling censorship.
4. concentrated solution filters: the cartridge filter of concentrated solution with 0.22 μ m film filtered, get filtering and concentrating liquid.
5. add the adjuvant lyophilizing: according to tiring of concentrated solution, the control finished product is above and tire in 70 units/more than the mg consumption of calculating gelatin, lactose than vigor 350 units/mg albumen.And respectively with the water for injection heating for dissolving and by 0.22 μ m membrane filtration, to be cooled to below 25 ℃, add the laggard freeze drying box lyophilizing of mix homogeneously in the filtering and concentrating liquid.The even packing 1～3L of every dish liquid is decided lyophilizing during lyophilizing on total amount of liquid.The freeze-dry process condition is: pre-freeze: products temperature-20 ℃--40 ℃, liquid measure in the optic disc is incubated 2-4 hour.Distillation: 10 ℃/hour of conduction oil programming rates, to oily temperature rise to 30 ℃, case vacuum is not more than 15Pa before the process control.Dry: as when products temperature reaches 23 ℃, to be incubated more than 4 hours until dry terminal point.
6. sieve, packing: finished product that lyophilizing is good is pulverized and is crossed behind 80 mesh sieves and the mix homogeneously with medicinal complex pocket packing, is loaded in aluminum listens, and seals.
The new technology of table 1 crude drug and original technology purity contrast table
Select five lot numbers
Conclusion: new solution purity is higher
Original technical scheme: the original method that adopts of inventor is by ion exchange chromatography single step purification kallidinogenase crude product, adopts 3-4 times of acetone precipitation, the kallidinogenase that purification obtains.
New technology standard: the scheme of employing new technology
The new technology of table 2 crude drug and original technology leading indicator contrast table
| Project | The new technology standard | Original technology |
| Purity | 85％ | 65％ |
| pH | 5.8～7.2 | 5.5～7.5 |
| Loss on drying | Be no more than 4.0% | Be no more than 5.0% |
| Tire (increasing item) | 55 units/mg | ------ |
| Than living | 350 units/mg albumen | 300 units/mg albumen |
Conclusion: novel technique is produced the every index of product and all is better than original technology
The different freeze temperature of table 3 is to the influence of purity and pH
| Products temperature | Purity | pH |
| -60℃ | 66％ | 8.4 |
| -50℃ | 71％ | 8.1 |
| -40℃ | 82％ | 7.2 |
| -30℃ | 84％ | 6.8 |
| -20℃ | 83％ | 5.9 |
| -10℃ | 76％ | 4.3 |
| -5℃ | 69％ | 4.2 |
Conclusion :-20 ℃----40 ℃ of purity are higher, and pH is more stable
Claims (8)
1. method for preparing kallidin proenzyme raw-material medicine is characterized in that may further comprise the steps:
1. preparation: the NH of preparation 0.15-0.5mol/L
4The NH of AC buffer and 0.1-0.5mol/L
4The AC buffer is adjusted to pH6.0, and surveys electric conductivity value;
2. chromatography:
2.1 balance: the chromatographic column of QAE-Sepharose Fast Flow ion column filler will be housed, use the NH of the 0.15-0.5mol/L of pH6.0
4AC buffer balance finishes until turnover solution pH value unanimity, the identical back balance of electric conductivity value;
2.2 sample introduction: transfer pH value and the balance liquid of kallidinogenase intermediate close, promptly 6.1～6.2, electric conductivity value is lower than the balance liquid electric conductivity value, promptly hangs down 0.2～0.3 * 10
4μ s/cm is according to every liter of filler maximum exchange amount 2,000 ten thousand units calculating applied sample amount and with the sample solution upper prop;
2.3 washing: the NH that uses the 0.15-0.5mol/L of pH6.0
4AC buffer flushing pillar is followed the tracks of with spectrophotometer simultaneously and is detected the absorption value of effluent at the 280nm place, washes to be lower than at 0.5 o'clock to eluate OD value and to stop;
2.4 eluting: the NH that uses the 0.1-0.5mol/L of pH6.0
4The AC buffer is followed the tracks of with spectrophotometer simultaneously and is detected its absorption value at the 253nm place with 500～600ml/min flow velocity eluting pillar, collects peak liquid;
3. ultrafiltration desalination: will collect liquid and add the purified water desalination and concentration by ultrafiltration, to exceeding the liquid electric conductivity value less than 0.1 * 10
4When μ s/cm is following, promptly stop to add purified water, continue to concentrate;
4. concentrated solution filters;
5. add the adjuvant lyophilizing;
6. sieve, packing.
2. the described method for preparing kallidin proenzyme raw-material medicine of claim 1, wherein used chromatographic column is ￠ 40cm during chromatography, volume is 50L.
3. the described method for preparing kallidin proenzyme raw-material medicine of claim 1 is that eluting is collected starting point when wherein tiring greater than 50 units/cm with eluent, and tiring less than 50 units/cm with eluent is that eluting is collected terminal point, and collecting peak liquid long-pending is 30～60L.
4. the described method for preparing kallidin proenzyme raw-material medicine of claim 1, wherein the molecular cut off during the ultrafiltration desalination is 10000dt.
5. the described method for preparing kallidin proenzyme raw-material medicine of claim 1, wherein in the step 3 when being concentrated into liquid volume 6～11L, close ultrafilter, purified water is towards film, tire when being lower than 50 units/ml to return pipe, concentrated solution, the sampling censorship.
6. the described method for preparing kallidin proenzyme raw-material medicine of claim 1 wherein in the step 4 is filtered the cartridge filter of concentrated solution with 0.22 μ m film, gets filtering and concentrating liquid.
7. the described method for preparing kallidin proenzyme raw-material medicine of claim 1, it is as follows wherein to add the freeze dried step of adjuvant: according to tiring of concentrated solution, the control finished product than vigor 350 units/mg albumen above with tire in 70 units/more than the mg, calculate the consumption of gelatin, lactose, and respectively with the water for injection heating for dissolving and by 0.22 μ m membrane filtration, to be cooled to below 25 ℃, add the laggard freeze drying box lyophilizing of mix homogeneously in the filtering and concentrating liquid; The even packing 1～3L of every dish liquid is decided on total amount of liquid during lyophilizing; The freeze-dry process condition is: pre-freeze: products temperature-20 ℃--40 ℃, liquid measure in the optic disc is incubated 2-4 hour.
8. the described method for preparing kallidin proenzyme raw-material medicine of claim 1, wherein that lyophilizing is good finished product are pulverized and are crossed behind 80 mesh sieves and the mix homogeneously with medicinal complex pocket packing, are loaded in aluminum listens, and seal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100982895A CN101073666B (en) | 2007-05-11 | 2007-05-11 | Method for producing kallidin proenzyme raw-material medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100982895A CN101073666B (en) | 2007-05-11 | 2007-05-11 | Method for producing kallidin proenzyme raw-material medicine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101073666A CN101073666A (en) | 2007-11-21 |
| CN101073666B true CN101073666B (en) | 2010-11-17 |
Family
ID=38975044
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100982895A Active CN101073666B (en) | 2007-05-11 | 2007-05-11 | Method for producing kallidin proenzyme raw-material medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101073666B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101914511B (en) * | 2010-04-27 | 2012-01-25 | 宁波林叶生物科技有限公司 | Method for preparing high-purity pancreatic kininogenase |
| CN105176953A (en) * | 2015-08-03 | 2015-12-23 | 常州千红生化制药股份有限公司 | Pancreatic kallidinogenase purifying process |
| CN113121675B (en) * | 2019-12-31 | 2023-03-31 | 翰宇药业（武汉）有限公司 | Salt conversion method of GLP-1 analogue |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1336434A (en) * | 2000-08-01 | 2002-02-20 | 上海惠海生化制品厂 | Prepn. and affinity chromatographic purification process of kallidinogen enzyme |
-
2007
- 2007-05-11 CN CN2007100982895A patent/CN101073666B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1336434A (en) * | 2000-08-01 | 2002-02-20 | 上海惠海生化制品厂 | Prepn. and affinity chromatographic purification process of kallidinogen enzyme |
Non-Patent Citations (3)
| Title |
|---|
| JP昭55-99191A 1980.07.28 |
| 周祖荫等.高纯度猪胰激肽释放酶的制备.中国生化药物杂志 1.1986,(1),6-11. |
| 周祖荫等.高纯度猪胰激肽释放酶的制备.中国生化药物杂志 1.1986,(1),6-11. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101073666A (en) | 2007-11-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102875669B (en) | Method for separating and extracting ovotransferrin | |
| CN103073638A (en) | Method for purifying ulinastatin via affinity chromatography | |
| CN103725664A (en) | Method for highly purifying kallikrein | |
| CN105648008A (en) | Feeding silk antibacterial peptide preparation and preparation method thereof | |
| CN101073666B (en) | Method for producing kallidin proenzyme raw-material medicine | |
| CN107011433B (en) | Extraction and purification method of nostoc sphaeroides biliprotein and purified phycocyanin | |
| CN107043431A (en) | The purification process of bacillary capsular polysaccharide | |
| CN104109204A (en) | Method for separating and purifying recombinant human lactoferrins from paddy rice seeds | |
| CN105287690A (en) | Bilberry extract and preparation method thereof | |
| CN108118044A (en) | A kind of method of separating-purifying egg white lysozyme | |
| CN101456898B (en) | Method for separating and purifying polypeptide by using hydrogen bond adsorption chromatogram | |
| CN109942723A (en) | A kind of separation method of polysaccharides | |
| CN1587275A (en) | Separating and purifying technology for micro algae phycoerythrin | |
| CN102787111A (en) | Preparation process for high-purity pancreatic kininogenase | |
| CN101544968B (en) | Industrial production method of earthworm fibrinolytic enzyme directly extracted by utilizing ion exchange resin | |
| CN108186790A (en) | A kind of glutinous rehmannia extract, preparation method and the application in terms of CIK cell in vitro proliferation is promoted | |
| CN110002962B (en) | Method for extracting alkyl resorcinol from wheat bran | |
| CN109734796B (en) | Process for separating albumin from haemolytic serum | |
| CN107200774A (en) | A kind of phycoerythrin of the extraction of nostoc phycobniliprotein, purification process and purifying | |
| CN114230689B (en) | Affinity resin for separating phycoerythrin, preparation method and application thereof | |
| CN118271424B (en) | Separation preparation method of fresh milk-grade high-purity alpha-lactalbumin | |
| CN1206240C (en) | High efficiency ionic exchange membrane chromatography decontamination vitelloenin technology | |
| CN114316088B (en) | Affinity resin, preparation method and application thereof in separation and purification of phycocyanin | |
| CN108690103B (en) | Method for preparing high-purity glucoraphanin extract by taking radish seeds as raw materials | |
| CN114933632B (en) | Method for preparing 5 kinds of ustilaginoidea virens toxins through simultaneous separation and purification |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |